TY - JOUR
T1 - Analysis of a rare progeria variant of Barrier-to-autointegration factor in Drosophila connects centromere function to tissue homeostasis
AU - Duan, Tingting
AU - Thyagarajan, Srikantha
AU - Amoiroglou, Anastasia
AU - Rogers, Gregory C.
AU - Geyer, Pamela K.
N1 - Funding Information:
We thank S. Cole Kitzman for technical assistance and other members of the Geyer laboratory for helpful discussions. We thank B. Mellone (U Connecticut) and N. Rusan (NIH) or generously supplying CENP-C and Centrosomin (Cnn) antibodies, respectively. We thank K. Furukawa (Niigata U, Japan) for generously supplying the baf mutant stock and Ting Xie (Hongkong University of Science and Technology, Hongkong) for generously supplying the chk2 mutant stock. This work was supported by the National Institute of Health GM087341 to P.K.G. Imaging was supported by the University of Iowa Central Microscopy Research Facility and use of the Zeiss LSM710 confocal microscope acquired via National Institutes of Health (NIH) funding (S10 RR025439-01). 1 KD
Funding Information:
We thank S. Cole Kitzman for technical assistance and other members of the Geyer laboratory for helpful discussions. We thank B. Mellone (U Connecticut) and N. Rusan (NIH) or generously supplying CENP-C and Centrosomin (Cnn) antibodies, respectively. We thank K. Furukawa (Niigata U, Japan) for generously supplying the baf1mutant stock and Ting Xie (Hongkong University of Science and Technology, Hongkong) for generously supplying the chk2KDmutant stock. This work was supported by the National Institute of Health GM087341 to P.K.G. Imaging was supported by the University of Iowa Central Microscopy Research Facility and use of the Zeiss LSM710 confocal microscope acquired via National Institutes of Health (NIH) funding (S10 RR025439-01).
Publisher Copyright:
© 2023, The Author(s).
PY - 2023/3
Y1 - 2023/3
N2 - Barrier-to-autointegration factor (BAF/BANF) is a nuclear lamina protein essential for nuclear integrity, chromatin structure, and genome stability. Whereas complete loss of BAF causes lethality in multiple organisms, the A12T missense mutation of the BANF1 gene in humans causes a premature aging syndrome, called Néstor-Guillermo Progeria Syndrome (NGPS). Here, we report the first in vivo animal investigation of progeroid BAF, using CRISPR editing to introduce the NGPS mutation into the endogenous Drosophila baf gene. Progeroid BAF adults are born at expected frequencies, demonstrating that this BAF variant retains some function. However, tissue homeostasis is affected, supported by studies of the ovary, a tissue that depends upon BAF for stem cell survival and continuous oocyte production. We find that progeroid BAF causes defects in germline stem cell mitosis that delay anaphase progression and compromise chromosome segregation. We link these defects to decreased recruitment of centromeric proteins of the kinetochore, indicating dysfunction of cenBAF, a localized pool of dephosphorylated BAF produced by Protein Phosphatase PP4. We show that DNA damage increases in progenitor germ cells, which causes germ cell death due to activation of the DNA damage transducer kinase Chk2. Mitotic defects appear widespread, as aberrant chromosome segregation and increased apoptosis occur in another tissue. Together, these data highlight the importance of BAF in establishing centromeric structures critical for mitosis. Further, these studies link defects in cenBAF function to activation of a checkpoint that depletes progenitor reserves critical for tissue homeostasis, aligning with phenotypes of NGPS patients.
AB - Barrier-to-autointegration factor (BAF/BANF) is a nuclear lamina protein essential for nuclear integrity, chromatin structure, and genome stability. Whereas complete loss of BAF causes lethality in multiple organisms, the A12T missense mutation of the BANF1 gene in humans causes a premature aging syndrome, called Néstor-Guillermo Progeria Syndrome (NGPS). Here, we report the first in vivo animal investigation of progeroid BAF, using CRISPR editing to introduce the NGPS mutation into the endogenous Drosophila baf gene. Progeroid BAF adults are born at expected frequencies, demonstrating that this BAF variant retains some function. However, tissue homeostasis is affected, supported by studies of the ovary, a tissue that depends upon BAF for stem cell survival and continuous oocyte production. We find that progeroid BAF causes defects in germline stem cell mitosis that delay anaphase progression and compromise chromosome segregation. We link these defects to decreased recruitment of centromeric proteins of the kinetochore, indicating dysfunction of cenBAF, a localized pool of dephosphorylated BAF produced by Protein Phosphatase PP4. We show that DNA damage increases in progenitor germ cells, which causes germ cell death due to activation of the DNA damage transducer kinase Chk2. Mitotic defects appear widespread, as aberrant chromosome segregation and increased apoptosis occur in another tissue. Together, these data highlight the importance of BAF in establishing centromeric structures critical for mitosis. Further, these studies link defects in cenBAF function to activation of a checkpoint that depletes progenitor reserves critical for tissue homeostasis, aligning with phenotypes of NGPS patients.
KW - Cell division
KW - Checkpoint kinase 2
KW - Chromosome segregation
KW - DNA damage response
KW - Disease variants
KW - Oogenesis
KW - Premature aging
KW - Tissue homeostasis
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UR - http://www.scopus.com/inward/citedby.url?scp=85149021109&partnerID=8YFLogxK
U2 - 10.1007/s00018-023-04721-y
DO - 10.1007/s00018-023-04721-y
M3 - Article
C2 - 36842139
AN - SCOPUS:85149021109
SN - 1420-682X
VL - 80
JO - Cellular and Molecular Life Sciences
JF - Cellular and Molecular Life Sciences
IS - 3
M1 - 73
ER -